Filter
Top Products
PA0027
Page 3
• Consult Federal, State or local regulations for disposal of any potentially toxic components.
• Minimize microbial contamination of reagents or an increase in non-specic staining may occur.
• Retrieval, incubation times or temperatures other than those specied may give erroneous results. Any such change must be
validated by the user.
Instructions for Use
Melanoma Marker (HMB45) primary antibody is recommended for use on an automated Bond system in combination with either
Bond Polymer Rene Detection (DS9800) or Bond Polymer Rene Red Detection (DS9390). The recommended staining protocols for
Melanoma Marker (HMB45) primary antibody are IHC Protocol F when using Bond Polymer Rene Detection and IHC Protocol J when
using Bond Polymer Rene Red Detection. Enzyme pretreatment is recommended using Bond Enzyme 1 for 5 minutes.
Results Expected
Normal Tissues
Melanoma Marker (HMB45) showed positive granular, cytoplasmic staining in melanocytes in the basal layer of epidermis. All other
normal tissues were negative. (Total number of cases stained = 99).
Tumor Tissues
Melanoma Marker (HMB45) stained 13/13 malignant melanomas. No staining was observed in a variety of other tumors. (Total number
of cases stained = 101).
Melanoma Marker (HMB45) is recommended for use as part of a panel of antibodies for the diagnosis of melanomas.
Product Specic Limitations
Melanoma Marker (HMB45) is recommended for use with either Bond Polymer Rene Detection or Bond Polymer Rene Red Detection
and Bond ancillary reagents. Users who deviate from recommended test procedures must accept responsibility for interpretation
of patient results under these circumstances. The protocol times may vary, due to variation in tissue xation and the effectiveness
of antigen enhancement, and must be determined empirically. Negative reagent controls should be used when optimizing retrieval
conditions and protocol times.
Troubleshooting
Refer to reference 3 for remedial action.
Contact your local distributor or the regional ofce of Leica Biosystems to report unusual staining.
Further Information
Further information on immunostaining with Bond reagents, under the headings Principle of the Procedure, Materials Required,
Specimen Preparation, Quality Control, Assay Verication, Interpretation of Staining, Key to Symbols on Labels, and General Limitations
can be found in “Using Bond Reagents” in your Bond user documentation.
Bibliography
1. Clinical Laboratory Improvement Amendments of 1988, Final Rule 57 FR 7163 February 28, 1992.
2. Villanova PA. National Committee for Clinical Laboratory Standards (NCCLS). Protection of laboratory workers from infectious
diseases transmitted by blood and tissue; proposed guideline. 1991; 7(9). Order code M29-P.
3. Bancroft JD and Stevens A. Theory and Practice of Histological Techniques. 4th Edition. Churchill Livingstone, New York. 1996.
4. Kapur RP, Bigler SA, Skelly M, et al. Anti-melanoma monoclonal antibody HMB45 identies an oncofetal glycoconjugate associated
with immature Melanoma Markers. The Journal of Histochemistry and Cytochemistry. 1992; 40(2):207-212.
5. Gown AM, Vogel AM, Hoak D, et al. Monoclonal antibodies specic for melanocytic tumours distinguish subpopulations of
melanocytes. American Journal of Pathology. 1986;123(2):195-203.
ProClin
™
950 is a trademark of Supelco, a part of Sigma-Aldrich Corporation.
Date of Issue
09 May 2013